Zedoary turmeric oil injection ameliorates lung inflammation via platelet factor 4 and regulates gut microbiota disorder in respiratory syncytial virus-infected young mice

Background Respiratory syncytial virus (RSV)-induced lung inflammation is one of the main causes of hospitalization and easily causes disruption of intestinal homeostasis in infants, thereby resulting in a negative impact on their development. However, the current clinical drugs are not satisfactory. Zedoary turmeric oil injection (ZTOI), a patented traditional Chinese medicine (TCM), has been used for clinical management of inflammatory diseases. However, its in vivo efficacy against RSV-induced lung inflammation and the underlying mechanism remain unclear. Purpose The present study was designed to confirm the in vivo efficacy of ZTOI against lung inflammation and intestinal disorders in RSV-infected young mice and to explore the potential mechanism. Study design and methods Lung inflammation was induced by RSV, and cytokine antibody arrays were used to clarify the effectiveness of ZTOI in RSV pneumonia. Subsequently, key therapeutic targets of ZTOI against RSV pneumonia were identified through multi-factor detection and further confirmed. The potential therapeutic material basis of ZTOI in target tissues was determined by non-target mass spectrometry. After confirming that the pharmacological substances of ZTOI can reach the intestine, we used 16S rRNA-sequencing technology to study the effect of ZTOI on the intestinal bacteria. Results In the RSV-induced mouse lung inflammation model, ZTOI significantly reduced the levels of serum myeloperoxidase, serum amyloid A, C-reactive protein, and thymic stromal lymphoprotein; inhibited the mRNA expression of IL-10 and IL-6; and decreased pathological changes in the lungs. Immunofluorescence and qPCR experiments showed that ZTOI reduced RSV load in the lungs. According to cytokine antibody arrays, platelet factor 4 (PF4), a weak chemotactic factor mainly synthesized by megakaryocytes, showed a concentration-dependent change in lung tissues affected by ZTOI, which could be the key target for ZTOI to exert anti-inflammatory effects. Additionally, sesquiterpenes were enriched in the lungs and intestines, thereby exerting anti-inflammatory and regulatory effects on gut microbiota. Conclusion ZTOI can protect from lung inflammation via PF4 and regulate gut microbiota disorder in RSV-infected young mice by sesquiterpenes, which provides reference for its clinical application in RSV-induced lung diseases. Supplementary Information The online version contains supplementary material available at 10.1186/s13020-024-00954-6.


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Figure S1 Representative histological images of hematoxylin and eosin-stained lung section (n=5).

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Figure S2 Changes of key inflammatory indicators in lung and serum (myeloperoxidase, MPO; serum amyloid A, SAA; C-reactive protein, CRP; thymic stromal lymphoprotein, TSLP; n = 6; data are presented as Means ± SD; ** P < 0.01, compared with the control group.

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Figure S3 Lung indexes evaluation.

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FigureS5RNA expression level of RSV, n > 6; *** P < 0.001, compared with the RSV group; ### P < 0.001, compared with the control group; data presented as mean ± standard error of mean)

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Figure S7 Forty cytokines and their distribution in chip.

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Figure S8 RNA expression level of PF4.

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Figure S9 Detail fragmentation pathway of curdione.

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Figure S10 Detail fragmentation pathway of germacrone.

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Figure S16 Ternary analysis of the fecal microorganisms composition in the control, RSV and ZTOI-H groups at genus level.

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Figure S17 Differential cytokines analysis of the RSV group compared to the control group .

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Figure S18 Viral protein interaction with cytokine and cytokine receptor pathway.

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Figure S19 TNF signaling pathway.

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Figure S20 Relationship of differential cytokines in the RSV group compared to the control group (Purple: cytokine enriched in TNF signaling pathway).

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Figure S21 Relationship of differential cytokines in the ZTOI group compared to the RSV group

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Figure S25 Original western blot figures of figure 5A.